Effects of human tissue kallikerin gene delivery on the proliferation of vascular smooth muscle cells
10.3760/cma.j.issn.0253-3758.2010.08.014
- VernacularTitle:人激肽释放酶基因转移对大鼠血管平滑肌细胞增殖的影响
- Author:
Peng-Li ZHU
1
;
Hui-Zhen YU
;
Liang-Di XIE
;
Jing-Ming RUAN
;
Chang-Sheng XU
;
Ti-Yuan LI
Author Information
1. 福建省立医院
- Keywords:
Blood vessels;
Myocytes,smooth muscle;
Cell proliferation;
Tissue kallikreins;
Gene transfer techniques
- From:
Chinese Journal of Cardiology
2010;38(8):739-744
- CountryChina
- Language:Chinese
-
Abstract:
Objective Tissue kallikrein cleaves kininogen substrate to produce vasoactive kinin peptides that have been implicated in the proliferation of vascular smooth muscle cells. We investigated the effects of adenovirus-mediated human tissue kallikerin(Ad-hKLK1)gene delivery on the proliferation of vascular smooth muscle cells of SHR(VSMCsSHR)induced by platelet derived growth factor-BB(PDGF-BB). Methods Primary VSMCsSHR were isolated and cultured from thoracic aorta of male SHR. The VSMCsSHR proliferation induced by PDGF-BB was accessed by cell counting and methyl thiazolyl tetrazoliuin(MTT). Western blot was used to determine the protein expression of hKLK1, the cycle-independent kinase inhibitors p27Kip1 and p21Cip1 . The mRNA expressions of bradykinin B1 receptor and B2 receptor were detected by RT-PCR in VSMCsSHR. Results Proliferation of VSMCsSHR induced by PDGF-BB was significantly inhibited post transfection of Ad-hKLK1(20 - 100 MOI)in a MOI-dependent manner. The peak inhibition titer of Ad-hKLK1 was 100 MOI with peak inhibition rate of 39.3%(cell counting, n = 3,P ＜0.01), 30.2%(MTT, n-3 ,P ＜ 0.01)and 36.4%(peak stunning rate of cell-cycle in phase G0/G1). The inhibitory effects of proliferation and cell-cycle caused by hKLK1 gene delivery could be abolished by Hoe140, a bradykinin B2 receptor antagonist. The protein expression of p27Kip1 and p21Cip1 increased significantly after the hKLK1 gene delivery, whereas Hoe140 nearly completely blocked these effects(n = 3,P ＜0.001 ,respectively). PDGF-BB also significantly upregulated the mRNA expression of B2 receptor but not B1 receptor in VSMCsSHR. Conclusion The hKLK1 gene delivery could inhibit PDGF-BB induced proliferation in VSMCsSHR through Bradykinin B2 receptor and up-regulate expression of p27Kip1 and p21Cip1.
